Interaction of DNA-Dependent Protein Kinase and Poly(ADP-Ribose) Polymerase with Radiation-Induced DNA Strand Breaks

Weinfeld, Michael; Chaudhry, M. Ahmad; D’Amours, Damien; Pelletier, Joëlle D.; Poirier, Guy G.; Povirk, Lawrence F.; Lees‐Miller, Susan P.

doi:10.2307/3579534

Cited by 54 publications

(43 citation statements)

References 37 publications

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“…4C, lanes 2-6, respectively). The concentration of DNA used was shown previously to give maximum activation of DNA-PK (29). No stimulation of ATM activity toward RPA heterotrimer was observed (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Purification and Characterization of ATM from Human Placenta

Chan¹,

Son²,

Block³

et al. 2000

Journal of Biological Chemistry

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115

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ATM is mutated in the human genetic disorder ataxia telangiectasia, which is characterized by ataxia, immune defects, and cancer predisposition. Cells that lack ATM exhibit delayed up-regulation of p53 in response to ionizing radiation. Serine 15 of p53 is phosphorylated in vivo in response to ionizing radiation, and antibodies to ATM immunoprecipitate a protein kinase activity that, in the presence of manganese, phosphorylates p53 at serine 15. Immunoprecipitates of ATM also phosphorylate PHAS-I in a manganese-dependent manner. Here we have purified ATM from human cells using nine chromatographic steps. Highly purified ATM phosphorylated PHAS-I, the 32-kDa subunit of RPA, serine 15 of p53, and Chk2 in vitro. The majority of the ATM phosphorylation sites in Chk2 were located in the amino-terminal 57 amino acids. In each case, phosphorylation was strictly dependent on manganese. ATM protein kinase activity was inhibited by wortmannin with an IC 50 of approximately 100 nM. Phosphorylation of RPA, but not p53, Chk2, or PHAS-I, was stimulated by DNA. The related protein, DNA-dependent protein kinase catalytic subunit, also phosphorylated PHAS-I, RPA, and Chk2 in the presence of manganese, suggesting that the requirement for manganese is a characteristic of this class of enzyme.

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Section: Resultsmentioning

confidence: 99%

“…Sonicated calf thymus DNA was prepared as described previously (28). Irradiated pUC18 plasmid DNA was prepared as described previously (29). Single-stranded M13 circular plasmid DNA was obtained from Life Technologies, Inc.…”

Section: Methodsmentioning

confidence: 99%

Purification and Characterization of ATM from Human Placenta

Chan¹,

Son²,

Block³

et al. 2000

Journal of Biological Chemistry

Self Cite

115

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“…TCR of DNA from xirradiated human cells has been demonstrated using an antibody to 5-bromo-2-deoxyuridine in repair patches, and some of these repair patches could have resulted from SSB repair (3)(4)(5). The reason that transcription-coupled repair may not be observed with SSBs although they block transcription may be due to both the abundance of repair proteins that process strand breaks, such as poly(ADP-ribose)polymerase, XRCC1, and polynucleotide kinase (47,48), and their demonstrated rapid global repair (49).…”

Section: Discussionmentioning

confidence: 99%

Single-Stranded Breaks in DNA but Not Oxidative DNA Base Damages Block Transcriptional Elongation by RNA Polymerase II in HeLa Cell Nuclear Extracts

Kathe¹,

Shen

Wallace

2004

Journal of Biological Chemistry

135

105

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Transcription and repair of many DNA helix-distorting lesions such as cyclobutane pyrimidine dimers have been shown to be coupled in cells across phyla from bacteria to humans. The signal for transcription-coupled repair appears to be a stalled transcription complex at the lesion site. To determine whether oxidative DNA lesions can block correctly initiated human RNA polymerase II, we examined the effect of site-specifically introduced oxidative damages on transcription in HeLa cell nuclear extracts. We found that transcription was blocked by single-stranded breaks, common oxidative DNA lesions, when present in the transcribed strand of the transcription template. Cyclobutane pyrimidine dimers, which have been previously shown to block transcription both in vitro and in vivo, also blocked transcription in the HeLa cell nuclear transcription assay. In contrast, the oxidative DNA base lesions, 8-oxoguanine, 5-hydroxycytosine, and thymine glycol did not inhibit transcription, although pausing was observed with the thymine glycol lesion. Thus, DNA strand breaks but not oxidative DNA base damages blocked transcription by RNA polymerase II.Transcription-coupled repair (TCR) 1 is a specialized form of DNA repair where damages are repaired preferentially in the transcribed strand of actively transcribed genes (for reviews, see Refs. 1 and 2). TCR was originally believed to be a subpathway of nucleotide excision repair; however, ionizing radiation damage (3), 5,6-dihydroxy-5,6-dihydrothymine (thymine glycol; Tg) (4, 5), and 7,8-dihydro-8-oxoguanine (8-oxoG) (5) are removed in a TCR-dependent manner from human cells that lack nucleotide excision repair. Since Tg and 8-oxoG are small nonbulky lesions that are repaired primarily by the base excision repair (BER) pathway (for reviews, see Refs. 6 -8), TCR of Tg and 8-oxoG in cells that lack nucleotide excision repair (5) links TCR to BER. TCR of 8-oxoG has also been shown to occur in nonreplicating Escherichia coli cells (9). TCR of oxidative damage does not appear to be universal, since in Chinese hamster ovary cells, TCR of oxidative damage produced by photosensitization and oxidizing agents is not observed in the Dhfr and cFos genes (10 -12). Furthermore, DNA strand breaks, oxidative lesions also repaired by BER, do not appear to be repaired by TCR in the Dhfr gene from Chinese hamster ovary cells (13) or human colon cancer cells (14). Interestingly, recent measurements of TCR of cyclobutane pyrimidine dimers in the Hprt gene, integrated at different sites in Chinese hamster ovary cell chromosomes, have suggested that preferential repair of actively transcribed genes, as well as preferential repair of damages in the transcribed strand, is significantly affected by genomic context (15).The proposed signal for TCR is an RNA polymerase transcription complex stalled at a lesion, which recruits the repair proteins to the damage site (16 -18); the ability of a lesion on the transcribed strand to block the RNA polymerase transcription complex has been assumed to be crucial for T...

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“…Studies with various oligomeric substrates suggest that activation involves localized denaturation of the extreme end of DNA and threading of a few bases of the single strands into defined channels (Hammarsten et al, 2000). Activation is relatively insensitive to structural modifications in these single strands (Weinfeld et al, 1997;Gu et al, 1998;Martensson and Hammarsten, 2002), suggesting that even severely damaged termini could still activate DNA-PK. Activated DNA-PK phosphorylates a variety of DNA-binding proteins (Anderson and Lees-Miller, 1992), including several with known or putative roles in end-joining such as KU (LeesMiller et al, 1990;Chan et al, 1999;Labhart, 1999), XRCC4 (Leber et al, 1998), WRN (Yannone et al, 2001) and the Artemis nuclease .…”

Section: Structural and Biochemical Properties Of End-joining Proteinsmentioning

confidence: 99%

Regulation and mechanisms of mammalian double-strand break repair

2003

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The double-strand break (DSB) is believed to be one of the most severe types of DNA damage, and if left unrepaired is lethal to the cell. Several different types of repair act on the DSB. The most important in mammalian cells are nonhomologous end-joining (NHEJ) and homologous recombination repair (HRR). NHEJ is the predominant type of DSB repair in mammalian cells, as opposed to lower eucaryotes, but HRR has recently been implicated in critical cell signaling and regulatory functions that are essential for cell viability. Whereas NHEJ repair appears constitutive, HRR is regulated by the cell cycle and inducible signal transduction pathways. More is known about the molecular details of NHEJ than HRR in mammalian cells. This review focuses on the mechanisms and regulation of DSB repair in mammalian cells, the signaling pathways that regulate these processes and the potential crosstalk between NHEJ and HRR, and between repair and other stress-induced pathways with emphasis on the regulatory circuitry associated with the ataxia telangiectasia mutated (ATM) protein.

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Interaction of DNA-Dependent Protein Kinase and Poly(ADP-Ribose) Polymerase with Radiation-Induced DNA Strand Breaks

Cited by 54 publications

References 37 publications

Purification and Characterization of ATM from Human Placenta

Purification and Characterization of ATM from Human Placenta

Single-Stranded Breaks in DNA but Not Oxidative DNA Base Damages Block Transcriptional Elongation by RNA Polymerase II in HeLa Cell Nuclear Extracts

Regulation and mechanisms of mammalian double-strand break repair

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